Abstract
Kinases are involved in a broad spectrum of cell behaviors. A single kinase can interact with different ligands each eliciting a specific cellular response. Dissecting downstream signaling pathways of kinases is a key step to understanding physiological and pathological cell process. However, directing kinase activity to specific substrates remains challenging. Here, we present a new tool to selectively activate a kinase in a specific protein complex in living cells. This technology uses a rapamycin-inducible kinase activation coupled to interaction with FKBP12-binding domain (FRB) tagged protein. Here, we demonstrate application of this method by targeting Src to either p130Cas or FAK and discriminating cell mophodynamic changes downstream each of these signaling complexes.
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Acknowledgments
We would like to thank Dr. Klaus Hahn for his support and guidance in development of this method. This work was supported by NIH R21 RCA159179A grant and Chicago Biomedical Consortium Pilot Grant to Dr. A. Karginov.
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Ray, AM., Klomp, J.E., Collins, K.B., Karginov, A.V. (2017). Dissecting Kinase Effector Signaling Using the RapRTAP Methodology. In: Tan, AC., Huang, P. (eds) Kinase Signaling Networks. Methods in Molecular Biology, vol 1636. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7154-1_2
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DOI: https://doi.org/10.1007/978-1-4939-7154-1_2
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7154-1
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